Team:Cambridge/Project

From 2012.igem.org

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(Overall project)
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== Project Details==
== Project Details==
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As stated above, the main aim of the project was to develop a bio-sensing standard to promote a platform for the development of novel biosensors that may work in a wealth of different ways but that can all be characterised and coupled to an output that is predictable, reliable and most importantly meaningful.
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=== Standardise Outputs ===
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=== Part 2 ===
 
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=== Biosensors ===
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=== The Experiments ===
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As the main crux of this project is a standardised output, we have aimed to develop several biosensors employing different mechanisms to prove the extended functionality of the final product. So far most of the biosensors in the registry use an inducible promoter to express their reporter protein. We have used some of these as a proof of the ability with which a sensor can be adapted for our output. However, we also explored another modality of biosensing in the form of riboswitches which could be the way of the future providing a more standard way of designing input circuits and hopefully a faster sensing method as the transcription step is not needed (as with inducible promoters).
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=== Instrumentation ===
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=== Part 3 ===
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We have constructed a mechanical rotary device that is turned by an arduino-controlled motor to 'sense' from 6 different cuvettes that can be placed in the device and then left for automated detection. The arduino is also connected to two light sensors, one supplied with a blue and the other with an orange filter,  the ratio of the light intensity at blue and orange frequencies can be measured at predefined time intervals.
 +
The hardware is coupled to a graphical user interface (GUI) that was designed using wxpython. Python is particularly useful as the communication with the arduino microcontroller is done using serial programming for which python has standard libraries. However, before any communication takes place between the user and the device, the arduino is loaded to perform the basic functions which are written in C++. The arduino and python were chosen for the ease of use and open platform. Also, the arduino is cheap and python is free!
== Results ==
== Results ==

Revision as of 16:19, 18 July 2012

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Contents

Overall project

Abstract

Previous iGEM teams have characterised an impressive array of inducible promoters, along with other elements of biosensing circuitry. But, to date, the output from each is not consistent and, in spite of the unifying biobrick standards used, do not necessarily couple together to make integrated test kits. The Cambridge iGEM 2012 team aim to take the true meaning of biobricks to heart, by creating an open and applied biosensor standard available for use by all subsequent teams, as well as, potentially, by industry and researchers in the field.

The biosensor aims to be modular in design, allowing the kits to be tailored to an individual's requirements, and to use light as an output to allow computer interfacing. We aim to use two luciferases, one to give a read-out of the input, and the other to act as a standard to allow fluctuations in colony size to be taken into account. Furthermore, we shall be using B. subtilis as our chassis, with the view to making the most of the spore forming capacity of bacteria to send out desiccated kits with long shelf lives.

  • Update - We have found a candidate for a new biobrick to be submitted to the registry of parts. Baker et al, in their paper entitled 'Widespread Genetic Switches and Toxicity Resistance Proteins for Fluoride', have identified a fluoride sensitive riboswitch which we, the Cambridge iGEM 2012 team, feel would be an excellent means to test, as proof of concept, our biosensor design.
  • Update2 - We have found another new sensor candidate, the Mg2+ riboswitch from Dann III et al's paper 'Structure and Mechanism of a Metal-Sensing Regulatory RNA'.

Implementation

Project Details

As stated above, the main aim of the project was to develop a bio-sensing standard to promote a platform for the development of novel biosensors that may work in a wealth of different ways but that can all be characterised and coupled to an output that is predictable, reliable and most importantly meaningful.


Standardise Outputs

Biosensors

As the main crux of this project is a standardised output, we have aimed to develop several biosensors employing different mechanisms to prove the extended functionality of the final product. So far most of the biosensors in the registry use an inducible promoter to express their reporter protein. We have used some of these as a proof of the ability with which a sensor can be adapted for our output. However, we also explored another modality of biosensing in the form of riboswitches which could be the way of the future providing a more standard way of designing input circuits and hopefully a faster sensing method as the transcription step is not needed (as with inducible promoters).


Instrumentation

We have constructed a mechanical rotary device that is turned by an arduino-controlled motor to 'sense' from 6 different cuvettes that can be placed in the device and then left for automated detection. The arduino is also connected to two light sensors, one supplied with a blue and the other with an orange filter, the ratio of the light intensity at blue and orange frequencies can be measured at predefined time intervals.

The hardware is coupled to a graphical user interface (GUI) that was designed using wxpython. Python is particularly useful as the communication with the arduino microcontroller is done using serial programming for which python has standard libraries. However, before any communication takes place between the user and the device, the arduino is loaded to perform the basic functions which are written in C++. The arduino and python were chosen for the ease of use and open platform. Also, the arduino is cheap and python is free!


Results